Our long-term objective is to improve human male reproductive health by designing, constructing, and marketing an automated microscopy system for aneuploidy scoring of human and rodent sperm for use in testing the effects of toxic agents and for research and clinical applications. The system will integrate the latest technologies in image analysis software, computer hardware, automated microscopes and associated fluorescence in situ hybridization (FISH) probes. We will use our existing FISH analysis system (called BDI-MultiFluorTM) as a basis for the proposed system development. Our specific aims in Phase I are to: 1) evaluate the existing system and characterize its performance for analyzing human sperm labeled with multiple chromosome-specific FISH probes; 2) optimize sample preparation methods; 3) adapt the existing software and develop new software to meet sperm aneuploidy requirements; and 4) specify additional system design parameters needed for accurate, high speed automated sperm aneuploidy scoring. The human sperm FISH test assay developed and validated at LLNL using visual microscope criteria will initially be used to develop the automated system; mouse and rat assays will be applied in Phase II. Factors that will drive the design of the system include not only technical issues such as accuracy and processing throughput, but also ease-of-use and cost effectiveness.